Advancement in CRISPR technology
An international team of researchers has made CRISPR technology more accessible and standardized by simplifying its complex implementation. The simpler, faster CRISPR, which is presented in the journal Nature Communications, offers a broad platform for off-the shelf genome engineering that may lower the barrier of entry for this powerful technology.
“CRISPR technologies can be programmed to target specific sequences of genetic code and to edit DNA at precise locations, thus allowing research scientists to permanently modify genes in living cells and model organisms to explore gene function in the laboratory, including genes associated with human disease,” said co-first author Dr. David Marciano, instructor in the Olivier Lichtarge laboratory at Baylor College of Medicine.
“These technologies permit a ribonucleoprotein complex to cleave DNA at a specific sequence that base-pairs with a guide RNA in the complex. Modularity of the nucleic acid/protein complex allows researchers to specify the guide RNA sequence to target nearly any sequence. This greatly improves researchers’ ability to edit DNA,” said co-first author Dr. Toon Swings, postdoctoral scientist in the Jan Michiels laboratory at VIB-KU Leuven Center for Microbiology.
However, this approach presents some challenges, such as constraints on the sequences that can be targeted, the possibility of off-target effects and the requirement of a unique guide RNA for each target gene. Marciano, Swings and their colleagues established an international collaboration that led to a simple solution that circumvents all these issues.
“Toon and I had a set of projects in which we had to construct many mutations and guide RNAs for different genes in the bacterium E. coli. We realized we wouldn’t need a new guide RNA for each gene if we just targeted a universal sequence found in gene knockout collections. The sequence we targeted is found in many genetic collections of medically important bacteria and is even in some fruit fly collections,” Marciano said.